RESUMEN
Macrophages play crucial roles in combatting infectious disease by promoting inflammation and phagocytosis. Angiopoietin-like protein 2 (ANGPTL2) is a secreted factor that induces tissue inflammation by attracting and activating macrophages to produce inflammatory cytokines in chronic inflammation-associated diseases such as obesity-associated metabolic syndrome, atherosclerosis, and rheumatoid arthritis. Here, we asked whether and how ANGPTL2 activates macrophages in the innate immune response. ANGPTL2 was predominantly expressed in proinflammatory mouse bone marrow-derived differentiated macrophages (GM-BMMs) following GM-CSF treatment relative to anti-inflammatory cells (M-BMMs) established by M-CSF treatment. Expression of the proinflammatory markers IL-1ß, IL-12p35, and IL-12p40 significantly decreased in GM-BMMs from Angptl2-deficient compared with wild-type (WT) mice, suggestive of attenuated proinflammatory activity. We also report that ANGPTL2 inflammatory signaling is transduced through integrin α5ß1 rather than through paired immunoglobulin-like receptor B. Interestingly, Angptl2-deficient mice were more susceptible to infection with Salmonella enterica serovar Typhimurium than were WT mice. Moreover, nitric oxide (NO) production by Angptl2-deficient GM-BMMs was significantly lower than in WT GM-BMMs. Collectively, our findings suggest that macrophage-derived ANGPTL2 promotes an innate immune response in those cells by enhancing proinflammatory activity and NO production required to fight infection.
Asunto(s)
Angiopoyetinas/inmunología , Predisposición Genética a la Enfermedad , Inmunidad Innata , Macrófagos/inmunología , Óxido Nítrico/inmunología , Infecciones por Salmonella/inmunología , Salmonella typhimurium/inmunología , Proteína 2 Similar a la Angiopoyetina , Proteínas Similares a la Angiopoyetina , Angiopoyetinas/genética , Animales , Femenino , Ratones , Ratones Noqueados , Óxido Nítrico/genética , Infecciones por Salmonella/genéticaRESUMEN
OBJECTIVE: The objective of this review is to explore the experiences and attitudes of physicians, nurses, and nursing assistants regarding task-shifting and task-sharing in hospitals. INTRODUCTION: Despite multiple health care professionals performing overlapping tasks, the need for effective task-shifting and task-sharing remains a concern. Understanding task-shifting and task-sharing experiences, as well as the attitudes of health care providers in hospitals, is essential for providing safe and patient-appropriate care with limited human resources. INCLUSION CRITERIA: Qualitative studies that examine the experiences and attitudes of physicians, nurses, and nursing assistants in hospitals regarding task-shifting and task-sharing will be included. The review will include physicians, advanced practice nurses who are nurse practitioners or clinical nurse specialists, registered nurses, and nursing assistants. Midwives, pharmacists, occupational therapists, physical therapists, and students will be excluded. METHODS: PubMed, MEDLINE, CINAHL, PsycINFO, Cochrane Database, and Web of Science will be searched as part of a 3-step search strategy. We will search for unpublished research and gray literature using Google Scholar and ProQuest Dissertations and Theses. Inclusion criteria will be studies published in English or Japanese from the time each database was established to the present. The methodological quality of all studies will be evaluated by screening against the inclusion criteria and by at least 2 critical evaluations using the standardized JBI checklist. Synthesized results will be pooled by meta-aggregation and published as a ConQual Summary of Findings. REVIEW REGISTRATION: PROSPERO CRD42023409612.
RESUMEN
Sphingomyelin synthase 1 (SMS1) catalyzes the conversion of ceramide to sphingomyelin. Here, we generated and analyzed SMS1-null mice. SMS1-null mice exhibited moderate neonatal lethality, reduced body weight, and loss of fat tissues mass, suggesting that they might have metabolic abnormality. Indeed, analysis on glucose metabolism revealed that they showed severe deficiencies in insulin secretion. Isolated mutant islets exhibited severely impaired ability to release insulin, dependent on glucose stimuli. Further analysis indicated that mitochondria in mutant islet cells cannot up-regulate ATP production in response to glucose. We also observed additional mitochondrial abnormalities, such as hyperpolarized membrane potential and increased levels of reactive oxygen species (ROS) in mutant islets. Finally, when SMS1-null mice were treated with the anti-oxidant N-acetyl cysteine, we observed partial recovery of insulin secretion, indicating that ROS overproduction underlies pancreatic ß-cell dysfunction in SMS1-null mice. Altogether, our data suggest that SMS1 is important for controlling ROS generation, and that SMS1 is required for normal mitochondrial function and insulin secretion in pancreatic ß-cells.
Asunto(s)
Insulina/metabolismo , Mitocondrias/patología , Especies Reactivas de Oxígeno/metabolismo , Transferasas (Grupos de Otros Fosfatos Sustitutos)/fisiología , Animales , Antioxidantes/farmacología , Glucosa/farmacología , Insulina/deficiencia , Secreción de Insulina , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patología , Ratones , Ratones Noqueados , Mitocondrias/fisiología , Mutación , Fenotipo , Transferasas (Grupos de Otros Fosfatos Sustitutos)/deficienciaRESUMEN
BACKGROUND: Autoimmune diseases are characterized by a breakdown of immunologic tolerance, and this breakdown can lead to life-threatening or lifelong disorders. Moreover; drugs that are used to treat these diseases are few in number and are associated with many serious adverse effects. METHODS: We used the rat insulin promoter-glycoprotein mouse model to analyze the role of tunicamycin in the process of autoimmune diabetes; the P14 mouse model to analyze the effect of tunicamycin on CD8(+) T cells; chop knockout mice to analyze the role of tunicamycin on an endoplasmic reticulum stress model; and fluorescence-activated cell sorting, quantitative real-time polymerase chain reaction, and histologic methods. RESULTS: We found that a single dose of tunicamycin reduced the activation and pancreatic infiltration of CD8(+) T cells. This activity delayed the incidence of virus-induced diabetes and improved survival rates. CONCLUSION: Tunicamycin may offer therapeutic opportunities for T cell-mediated autoimmune diseases such as diabetes.
Asunto(s)
Linfocitos T CD8-positivos/efectos de los fármacos , Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Tunicamicina/uso terapéutico , Animales , Apoptosis , Enfermedades Autoinmunes/tratamiento farmacológico , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Enfermedades Autoinmunes/virología , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Proliferación Celular/efectos de los fármacos , Citocinas/efectos de los fármacos , Citocinas/metabolismo , Diabetes Mellitus Experimental/inmunología , Diabetes Mellitus Experimental/patología , Diabetes Mellitus Experimental/virología , Estrés del Retículo Endoplásmico , Citometría de Flujo , Glicoproteínas/genética , Glicoproteínas/inmunología , Glicoproteínas/metabolismo , Hipoglucemiantes/administración & dosificación , Inmunosupresores/inmunología , Inmunosupresores/uso terapéutico , Activación de Linfocitos/efectos de los fármacos , Virus de la Coriomeningitis Linfocítica/química , Virus de la Coriomeningitis Linfocítica/inmunología , Virus de la Coriomeningitis Linfocítica/patogenicidad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Páncreas/metabolismo , Páncreas/patología , Regiones Promotoras Genéticas , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción CHOP/genética , Factor de Transcripción CHOP/metabolismoRESUMEN
OBJECTIVE: To investigate whether and how the endoplasmic reticulum (ER) stress-induced, CCAAT/enhancer-binding protein-homologous protein (CHOP)-mediated pathway regulates myocardial ischemia/reperfusion injury. METHODS AND RESULTS: Wild-type and chop-deficient mice underwent 50 minutes of left coronary artery occlusion followed by reperfusion. Expression of chop and spliced x-box binding protein-1 (sxbp1) mRNA was rapidly and significantly increased in reperfused myocardium of wild-type mice. chop-deficient mice exhibited markedly reduced injury size after reperfusion compared with wild-type mice, accompanied by a decreasing number of terminal deoxynucleotidyl transferase dUTP nick-end labeling-positive cardiomyocytes. Interestingly, myocardial inflammation, as assessed by expression of inflammatory cytokines and chemokines and numbers of infiltrated inflammatory cells, was also attenuated in chop-deficient mice. Moreover, expression of interleukin-6 mRNA in response to lipopolysaccharide was enhanced by simultaneous stimulation with thapsigargin, a potent ER stressor, in wild-type cardiomyocytes but not in chop-deficient cardiomyocytes. Finally, we found that superoxide was produced in reperfused myocardium and that intravenous administration of edaravone, a free radical scavenger, immediately before reperfusion significantly suppressed the superoxide overproduction and subsequent expression of sxbp1 and chop mRNA, followed by reduced injury size in wild-type mice. CONCLUSIONS: The ER stress-induced, CHOP-mediated pathway, which is activated in part by superoxide overproduction after reperfusion, exacerbates myocardial ischemia/reperfusion injury by inducing cardiomyocyte apoptosis and myocardial inflammation.
Asunto(s)
Apoptosis , Retículo Endoplásmico/metabolismo , Inflamación/prevención & control , Daño por Reperfusión Miocárdica/prevención & control , Miocardio/metabolismo , Factor de Transcripción CHOP/deficiencia , Animales , Antipirina/análogos & derivados , Antipirina/farmacología , Apoptosis/efectos de los fármacos , Células Cultivadas , Quimiocinas/metabolismo , Citocinas/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Modelos Animales de Enfermedad , Edaravona , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/inmunología , Depuradores de Radicales Libres/farmacología , Regulación de la Expresión Génica , Inflamación/genética , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Mediadores de Inflamación/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Daño por Reperfusión Miocárdica/genética , Daño por Reperfusión Miocárdica/inmunología , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Miocardio/inmunología , Miocardio/patología , Empalme del ARN , ARN Mensajero/metabolismo , Factores de Transcripción del Factor Regulador X , Estrés Fisiológico , Superóxidos/metabolismo , Tapsigargina/farmacología , Factores de Tiempo , Factor de Transcripción CHOP/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Duchenne muscular dystrophy (DMD) is the most common, lethal, muscle-wasting disease of childhood. Previous investigations have shown that muscle macrophages may play an important role in promoting the pathology in the mdx mouse model of DMD. In the present study, we investigate the mechanism through which macrophages promote mdx dystrophy and assess whether the phenotype of the macrophages changes between the stage of peak muscle necrosis (4 weeks of age) and muscle regeneration (12 weeks). We find that 4-week-old mdx muscles contain a population of pro-inflammatory, classically activated M1 macrophages that lyse muscle in vitro by NO-mediated mechanisms. Genetic ablation of the iNOS gene in mdx mice also significantly reduces muscle membrane lysis in 4-week-old mdx mice in vivo. However, 4-week mdx muscles also contain a population of alternatively activated, M2a macrophages that express arginase. In vitro assays show that M2a macrophages reduce lysis of muscle cells by M1 macrophages through the competition of arginase in M2a cells with iNOS in M1 cells for their common, enzymatic substrate, arginine. During the transition from the acute peak of mdx pathology to the regenerative stage, expression of IL-4 and IL-10 increases, either of which can deactivate the M1 phenotype and promote activation of a CD163+, M2c phenotype that can increase tissue repair. Our findings further show that IL-10 stimulation of macrophages activates their ability to promote satellite cell proliferation. Deactivation of the M1 phenotype is also associated with a reduced expression of iNOS, IL-6, MCP-1 and IP-10. Thus, these results show that distinct subpopulations of macrophages can promote muscle injury or repair in muscular dystrophy, and that therapeutic interventions that affect the balance between M1 and M2 macrophage populations may influence the course of muscular dystrophy.
Asunto(s)
Arginina/metabolismo , Macrófagos/inmunología , Músculo Esquelético/patología , Distrofia Muscular de Duchenne/inmunología , Animales , Proliferación Celular , Células Cultivadas , Citocinas/inmunología , Humanos , Activación de Macrófagos , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Músculo Esquelético/inmunología , Músculo Esquelético/metabolismo , Distrofia Muscular Animal/genética , Distrofia Muscular Animal/inmunología , Distrofia Muscular Animal/metabolismo , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenotipo , Índice de Severidad de la EnfermedadRESUMEN
Endoplasmic reticulum (ER) stress has been shown to participate in many disease pathologies. Although recent reports have demonstrated that ER stress in chondrocytes is present in human osteoarthritis (OA), its role in the pathology of cartilage degeneration, such as chondrocyte apoptosis, remains unclear. In the present study, we investigated the expression of phosphorylated PERK (pPERK), ubiquitin (Ub), GRP78, CHOP, phosphorylated JNK (pJNK) and cleaved caspase-3 (C-CASP3) and the mRNA splicing of XBP1 (XBP1 splicing) in human OA cartilage by immunohistochemistry and RT-PCR. Additionally, human chondrocytes were treated with several concentrations of tunicamycin, an ER stress inducer, to assess the impact of ER stress on the mRNA expression of CHOP, XBP1 splicing and apoptosis, as determined by real-time PCR, RT-PCR and ELISA analyses respectively. In human OA cartilage, the number of chondrocytes expressing pPERK, Ub, CHOP and pJNK positively correlated with cartilage degeneration and the number of C-CASP3-positive chondrocytes. XBP1 splicing and GRP78 expression in severe OA containing the greatest number of C-CASP3-positive chondrocytes were similar to the levels in mild OA, however, XBP1 splicing was higher in moderate OA than in mild and severe OA. Tunicamycin dose dependently increased CHOP expression and apoptosis of cultured chondrocytes. Although tunicamycin upregulated XBP1 splicing in cultured chondrocytes, its impact on XBP1 splicing was weakened at higher concentrations. In conclusion, the present results indicate that ER stress may contribute to chondrocyte apoptosis along with OA progression, which was closely associated with an enhanced apoptotic response and a reduced protective response by the cells.
Asunto(s)
Apoptosis/fisiología , Cartílago/patología , Condrocitos/patología , Retículo Endoplásmico/fisiología , Osteoartritis de la Rodilla/patología , Estrés Fisiológico/fisiología , Anciano , Cartílago/metabolismo , Cartílago/fisiopatología , Células Cultivadas , Condrocitos/metabolismo , Chaperón BiP del Retículo Endoplásmico , Proteínas de la Matriz Extracelular/metabolismo , Femenino , Proteínas de Choque Térmico/metabolismo , Humanos , MAP Quinasa Quinasa 4/metabolismo , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/metabolismo , Osteoartritis de la Rodilla/fisiopatología , Estudios Retrospectivos , Factor de Transcripción CHOP/metabolismo , Ubiquitina/metabolismo , eIF-2 Quinasa/metabolismoRESUMEN
OBJECTIVE: To elucidate whether and how the endoplasmic reticulum (ER) stress-C/EBP homologous protein (CHOP) pathway in macrophages is involved in the rupture of atherosclerotic plaques. METHODS AND RESULTS: Increases in macrophage-derived foam cell death in coronary atherosclerotic plaques cause the plaque to become vulnerable, thus resulting in acute coronary syndrome. The ER stress-CHOP/growth arrest and DNA damage-inducible gene-153 (GADD153) pathway is induced in the macrophage-derived cells in atherosclerotic lesions and is involved in plaque formation. However, the role of CHOP in the final stage of atherosclerosis has not been fully elucidated. Many CHOP-expressing macrophages showed apoptosis in advanced ruptured atherosclerotic lesions in wild-type mice, whereas few apoptotic cells were observed in Chop(-/-) mice. The rupture of atherosclerotic plaques was significantly reduced in high cholesterol-fed Chop(-/-)/Apoe(-/-) mice compared with Chop(+/+)/Apoe(-/-) mice. Furthermore, using mice that underwent bone marrow transplantation, we showed that expression of CHOP in macrophages significantly contributes to the formation of ruptures. By using primary cultured macrophages, we further showed that unesterified free cholesterol derived from incorporated denatured low-density lipoprotein was accumulated in the ER and induced ER stress-mediated apoptosis in a CHOP-Bcl2-associated X protein (Bax) pathway-dependent manner. CONCLUSIONS: The ER stress-CHOP-Bax-mediated apoptosis in macrophages contributes to the instability of atherosclerotic plaques.
Asunto(s)
Aterosclerosis/patología , Aterosclerosis/fisiopatología , Retículo Endoplásmico/fisiología , Macrófagos/patología , Macrófagos/fisiología , Factor de Transcripción CHOP/fisiología , Síndrome Coronario Agudo/etiología , Síndrome Coronario Agudo/patología , Síndrome Coronario Agudo/fisiopatología , Animales , Apolipoproteínas E/deficiencia , Apolipoproteínas E/genética , Apoptosis/fisiología , Trasplante de Médula Ósea , Células Cultivadas , Colesterol/metabolismo , Células Espumosas/patología , Células Espumosas/fisiología , Expresión Génica , Técnicas In Vitro , Macrófagos Peritoneales/patología , Macrófagos Peritoneales/fisiología , Masculino , Ratones , Ratones Noqueados , Transducción de Señal , Estrés Fisiológico , Factor de Transcripción CHOP/deficiencia , Factor de Transcripción CHOP/genética , Proteína X Asociada a bcl-2/fisiologíaRESUMEN
Although recent reports suggest that the endoplasmic reticulum (ER) stress response is induced in association with the development of inflammatory bowel disease, its role in the pathogenesis of inflammatory bowel disease remains unclear. The CCAAT/enhancer-binding protein (C/EBP) homologous protein (CHOP) is a transcription factor that is involved in the ER stress response, especially ER stress-induced apoptosis. In this study, we found that experimental colitis was ameliorated in CHOP-null mice, suggesting that CHOP exacerbates the development of colitis. The mRNA expression of Mac-1 (CD11b, a positive regulator of macrophage infiltration), Ero-1alpha, and Caspase-11 (a positive regulator of interleukin-1beta production) in the intestine was induced with the development of colitis, and this induction was suppressed in CHOP-null mice. ERO-1alpha is involved in the production of reactive oxygen species (ROS); an increase in ROS production, which is associated with the development of colitis in the intestine, was suppressed in CHOP-null mice. A greater number of apoptotic cells in the intestinal mucosa of wild-type mice were observed to accompany the development of colitis compared with CHOP-null mice, suggesting that up-regulation of CHOP expression exacerbates the development of colitis. Furthermore, this CHOP activity appears to involve various stimulatory mechanisms, such as macrophage infiltration via the induction of Mac-1, ROS production via the induction of ERO-1alpha, interleukin-1beta production via the induction of Caspase-11, and intestinal mucosal cell apoptosis.
Asunto(s)
Colitis/etiología , Colitis/patología , Retículo Endoplásmico/metabolismo , Estrés Oxidativo , Especies Reactivas de Oxígeno/metabolismo , Factor de Transcripción CHOP/fisiología , Animales , Apoptosis/fisiología , Caspasas/genética , Caspasas/metabolismo , Caspasas Iniciadoras , Colitis/inducido químicamente , Immunoblotting , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Mucosa Intestinal/citología , Mucosa Intestinal/metabolismo , Peroxidación de Lípido , Antígeno de Macrófago-1/genética , Antígeno de Macrófago-1/metabolismo , Macrófagos/citología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Peroxidasa/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
Differentiation of committed osteoblasts is controlled by complex activities involving signal transduction and gene expression, and Runx2 and Osterix function as master regulators for this process. Recently, CCAAT/enhancer-binding proteins (C/EBPs) have been reported to regulate osteogenesis in addition to adipogenesis. However, the roles of C/EBP transcription factors in the control of osteoblast differentiation have yet to be fully elucidated. Here we show that C/EBP homologous protein (CHOP; also known as C/EBPzeta) is expressed in bone as well as in mesenchymal progenitors and primary osteoblasts. Overexpression of CHOP reduces alkaline phosphatase activity in primary osteoblasts and suppresses the formation of calcified bone nodules. CHOP-deficient osteoblasts differentiate more strongly than their wild-type counterparts, suggesting that endogenous CHOP plays an important role in the inhibition of osteoblast differentiation. Furthermore, endogenous CHOP induces differentiation of calvarial osteoblasts upon bone morphogenetic protein (BMP) treatment. CHOP forms heterodimers with C/EBPbeta and inhibits the DNA-binding activity as well as Runx2-binding activity of C/EBPbeta, leading to inhibition of osteocalcin gene transcription. These findings indicate that CHOP acts as a dominant-negative inhibitor of C/EBPbeta and prevents osteoblast differentiation but promotes BMP signaling in a cell-type-dependent manner. Thus, endogenous CHOP may have dual roles in regulating osteoblast differentiation and bone formation.
Asunto(s)
Adipogénesis , Osteoblastos/citología , Factor de Transcripción CHOP/metabolismo , Células 3T3-L1 , Animales , Proteínas Morfogenéticas Óseas/farmacología , Proteína beta Potenciadora de Unión a CCAAT/metabolismo , Células COS , Células Cultivadas , Chlorocebus aethiops , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Regulación hacia Abajo/genética , Perfilación de la Expresión Génica , Ratones , Modelos Biológicos , Osteocalcina/genética , Osteogénesis/efectos de los fármacos , Regiones Promotoras Genéticas/genética , Unión Proteica , Factor de Transcripción CHOP/genéticaRESUMEN
C/EBP homologous protein (CHOP) is one of the main mediating factors in the ER stress pathway. To elucidate the role of the ER stress-CHOP pathway in experimental pancreatitis, wild-type (Chop(+/+)) and Chop deficient (Chop(-/-)) mice were administered cerulein, a cholecystokinin analogue, or both cerulein and lipopolysaccharide (LPS). In cerulein-induced acute pancreatitis, ER stress, serum amylase elevation and histological interstitial edema were induced. However, there was no remarkable activation downstream of the CHOP pathway regardless of the presence or absence of CHOP. Whereas, in the cerulein and LPS model, inflammation-associated caspases (caspase-11, caspase-1) and IL-1beta, but not apoptosis-associated caspases, were activated. In Chop(-/-) mice, the expression levels of these mediators returned to basal levels resulting in a milder pancreatitis and decreased serum amylase level. These results indicated that the ER stress-CHOP pathway has a pivotal role in the acceleration of pancreatitis through the induction of inflammation-associated caspases and IL-1beta.
Asunto(s)
Pancreatitis/metabolismo , Pancreatitis/patología , Factor de Transcripción CHOP/metabolismo , Amilasas/metabolismo , Animales , Apoptosis/fisiología , Secuencia de Bases , Western Blotting , Ceruletida/administración & dosificación , Modelos Animales de Enfermedad , Inflamación/inducido químicamente , Inflamación/metabolismo , Inflamación/patología , Interleucina-1beta/metabolismo , Lipopolisacáridos/administración & dosificación , Ratones , Ratones Endogámicos C57BL , Pancreatitis/inducido químicamenteRESUMEN
BACKGROUND/AIM: Oxidative stress is known to be enhanced in hemodialysis patients, and one of its useful markers is plasma copper/zinc superoxide dismutase (Cu/Zn-SOD). The increase in plasma Cu/Zn-SOD can be inhibited by orally administered lipid-soluble vitamin E. We examined the antioxidative effects of water-soluble vitamin C administered orally on Cu/Zn-SOD levels in hemodialysis patients. METHODS: Vitamin C was orally administered to 16 maintenance hemodialysis patients before each dialysis session. Doses were increased from 200 to 1,000 mg over 3 months. The levels of plasma vitamin C and Cu/Zn-SOD and its mRNA expression in leukocytes were determined 1, 2, and 3 months after the start of vitamin C administration. Furthermore, the levels of oxidized and reduced forms of plasma vitamin C were determined before the start of vitamin C administration and before and after dialysis at 1,000-mg vitamin C doses. RESULTS: Following oral administration, the plasma levels of vitamin C and its oxidized form were increased. However, significant changes in plasma Cu/Zn-SOD or its mRNA expression in leukocytes were not observed. CONCLUSION: In maintenance hemodialysis patients, vitamin C administration resulted in a significant increase in the postdialysis level of the oxidized form of vitamin C, which suggested an increase in antioxidant effect. However, water-soluble vitamin C did not significantly suppress Cu/Zn-SOD expression enhancement.
Asunto(s)
Ácido Ascórbico/administración & dosificación , Ácido Ascórbico/sangre , Fallo Renal Crónico/sangre , Fallo Renal Crónico/rehabilitación , Diálisis Renal , Superóxido Dismutasa/sangre , Administración Oral , Biomarcadores/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Oxidación-Reducción , Estrés OxidativoRESUMEN
Overload of pancreatic beta cells in conditions such as hyperglycemia, obesity, and long-term treatment with sulfonylureas leads to beta cell exhaustion and type 2 diabetes. Because beta cell mass declines under these conditions, apparently as a result of apoptosis, we speculated that overload kills beta cells as a result of endoplasmic reticulum (ER) stress. The Akita mouse, which carries a conformation-altering missense mutation (Cys96Tyr) in Insulin 2, likewise exhibits hyperglycemia and a reduced beta cell mass. In the development of diabetes in Akita mice, mRNAs for the ER chaperone Bip and the ER stress-associated apoptosis factor Chop were induced in the pancreas. Overexpression of the mutant insulin in mouse MIN6 beta cells induced Chop expression and led to apoptosis. Targeted disruption of the Chop gene delayed the onset of diabetes in heterozygous Akita mice by 8-10 weeks. We conclude that ER overload in beta cells causes ER stress and leads to apoptosis via Chop induction. Our findings suggest a new therapeutic approach for preventing the onset of diabetes by inhibiting Chop induction or by increasing chaperone capacity in the ER.
Asunto(s)
Proteínas Potenciadoras de Unión a CCAAT/deficiencia , Proteínas Potenciadoras de Unión a CCAAT/genética , Diabetes Mellitus Tipo 2/prevención & control , Retículo Endoplásmico/metabolismo , Proteínas de Choque Térmico , Factores de Transcripción/deficiencia , Factores de Transcripción/genética , Animales , Apoptosis , Proteínas Portadoras/genética , Diabetes Mellitus Tipo 2/etiología , Diabetes Mellitus Tipo 2/genética , Chaperón BiP del Retículo Endoplásmico , Insulina/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Mutantes , Chaperonas Moleculares/genética , Mutación Missense , Fenotipo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Estrés Fisiológico/genética , Estrés Fisiológico/metabolismo , Factor de Transcripción CHOPRESUMEN
Nitric oxide (NO) is a multifunctional biomolecule involved in a variety of physiological and pathological processes, including regulation of blood vessel dilatation and anti-arteriosclerotic effects. However, a large amount of NO is toxic to the host and causes several diseases such as apoptosis, septic shock, and diabetes mellitus. Inducible-form NO synthase is induced in inflammatory diseases, including insulitis and arteriosclerosis. Endoplasmic reticulum (ER) stress pathway was first identified as a cellular response pathway induced by the accumulation of unfolded proteins in ER to preserve ER functions. Later it was found that ER stress pathway is also activated by various cellular stresses to protect cells, but when stresses are severe, apoptosis is induced to remove damaged cells. It is reported that NO and reactive oxygen species disturb ER functions, then ER stress-mediated apoptosis pathway is activated. CHOP/GADD153, which belongs to C/EBP transcription factor family, is induced in this process and mediates apoptosis. ER stress pathway induced by NO can be involved in the pathogenesis of various vascular diseases.
Asunto(s)
Retículo Endoplásmico/metabolismo , Óxido Nítrico/metabolismo , Estrés Fisiológico/etiología , Estrés Fisiológico/fisiopatología , Animales , Apoptosis , HumanosRESUMEN
Neurotrophic factors are essential for the survival of neurons. We found that the endoplasmic reticulum (ER) stress-C/EBP homologues protein (CHOP) pathway to be activated during neurotrophic factor deprivation-induced apoptosis in PC12 neuronal cells and in primary cultured neurons, and this apoptosis was suppressed in the neurons from chop(-/-) mice. In addition, we found that CHOP is expressed in the subventricular zone (SVZ) and striatum of the young adult mouse brain. The number of apoptotic cells in the SVZ decreased in chop(-/-) mice. These results indicate that the ER stress-CHOP pathway plays a role in neuronal apoptosis during the development of the brain.
Asunto(s)
Apoptosis , Cuerpo Estriado/metabolismo , Factores de Crecimiento Nervioso/fisiología , Neuronas/metabolismo , Factor de Transcripción CHOP/fisiología , Animales , Secuencia de Bases , Células Cultivadas , Cuerpo Estriado/citología , Cartilla de ADN , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Ratones , Neuronas/citología , Células PC12 , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The endoplasmic reticulum (ER) is an organelle in which proteins are modified. When unfolded proteins accumulate in the ER under various stresses, ER stress (ERS) pathways, including the induction of chaperones, are activated to protect the cell. However, when ERS is excessive, the cell undergoes apoptosis. This study investigated ERS in multiple myeloma cells (MMCs) because they contain a well-developed ER due to M-protein production. The myeloma cell line 12-PE underwent apoptosis via caspase-3 after treatment with thapsigargin (thap), an ERS inducer, while another cell line, U266, did not. To understand the mechanism regulating this heterogeneity, the induction of chaperones by thap was analysed. Chaperones were up-regulated in U266 cells but down-regulated in 12-PE cells, suggesting that chaperones contribute to cell survival under ERS. Analysis of XBP-1, a transcriptional inducer of chaperones, in freshly isolated MMCs from 22 myeloma cases revealed 10 cases with active XBP-1, who also showed significantly poorer survival (p < 0.05), suggesting that chaperone expression protects MMCs from apoptosis, thereby allowing tumor cell expansion. These results suggest that MMCs are subjected to ERS under certain circumstances and that chaperones are induced to protect the cells against such ERS. Inhibition of chaperones could be a new target for myeloma therapy.
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Resistencia a Antineoplásicos , Retículo Endoplásmico/metabolismo , Mieloma Múltiple/metabolismo , Estrés Oxidativo , Adulto , Anciano , Anciano de 80 o más Años , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Proteínas de Unión al ADN/efectos de los fármacos , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Retículo Endoplásmico/efectos de los fármacos , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Chaperonas Moleculares/metabolismo , Mieloma Múltiple/genética , Mieloma Múltiple/patología , Proteínas Nucleares/efectos de los fármacos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Estrés Oxidativo/efectos de los fármacos , Pronóstico , ARN Mensajero/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de Transcripción del Factor Regulador X , Estudios Retrospectivos , Análisis de Supervivencia , Tapsigargina/farmacología , Factores de Transcripción , Proteína 1 de Unión a la X-BoxRESUMEN
INTRODUCTION: The role of macrophages in tumor progression has generated contradictory evidence. We had previously demonstrated the ability of peritoneal macrophages from LMM3 murine mammary adenocarcinoma-bearing mice (TMps) to increase the angiogenicity of LMM3 tumor cells, mainly through polyamine synthesis. Here we investigate the ability of the parasympathetic nervous system to modulate angiogenesis induced by TMps through the activation of the muscarinic acetylcholine receptor (mAchR). METHODS: Peritoneal macrophages from female BALB/c mice bearing a 7-day LMM3 tumor were inoculated intradermally (3 x 10(5) cells per site) into syngeneic mice. Before inoculation, TMps were stimulated with the muscarinic agonist carbachol in the absence or presence of different muscarinic antagonists or enzyme inhibitors. Angiogenesis was evaluated by counting vessels per square millimeter of skin. The expression of mAchR, arginase and cyclo-oxygenase (COX) isoforms was analyzed by Western blotting. Arginase and COX activities were evaluated by urea and prostaglandin E2 (PGE2) production, respectively. RESULTS: TMps, which stimulate neovascularization, express functional mAchR, because carbachol-treated TMps potently increased new blood vessels formation. This response was completely blocked by preincubating TMps with pirenzepine and 4-diphenylacetoxy-N-methylpiperidine (4-DAMP), M1 and M3 receptor antagonists, and partly by the M2 receptor antagonist methoctramine. M1 receptor activation by carbachol in TMps triggers neovascularization through arginase products because Nomega-hydroxy-L-arginine reversed the agonist action. Preincubation of TMps with methoctramine partly prevented carbachol-stimulated urea formation. In addition, COX-derived liberation of PGE2 is responsible for the promotion of TMps angiogenic activity by M3 receptor. We also detected a higher expression of vascular endothelial growth factor (VEGF) in TMps than in macrophages from normal mice. Carbachol significantly increased VEGF expression in TMps, and this effect was totally reversed by methoctramine and pirenzepine. Arginase and COX inhibitors partly decreased VEGF derived from TMps. CONCLUSION: TMps themselves induce a potent angiogenic response that is augmented by carbachol action. mAchR activation triggers arginine metabolism, PGE2 synthesis and VEGF production, promoting neovascularization.
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Adenocarcinoma/fisiopatología , Neoplasias Mamarias Animales/fisiopatología , Neovascularización Patológica , Receptores Muscarínicos/fisiología , Animales , Arginasa/biosíntesis , Arginasa/metabolismo , Western Blotting , Femenino , Perfilación de la Expresión Génica , Macrófagos Peritoneales/inmunología , Ratones , Ratones Endogámicos BALB C , Prostaglandina-Endoperóxido Sintasas/biosíntesis , Prostaglandina-Endoperóxido Sintasas/metabolismo , Isoformas de Proteínas , Receptores Muscarínicos/biosíntesisRESUMEN
CHOP is a C/EBP family transcription factor involved in endoplasmic reticulum (ER) stress-mediated apoptosis. To determine if the ER stress pathway is involved in the pathogenesis of LPS-treated mouse lung injury, mice were given lipopolysaccharide (LPS) intraperitoneally. The mRNAs for activating transcription factor (ATF) 4 and X-box binding protein (XBP) 1, transcriptional activators of the CHOP gene, and that for CHOP were induced by or after the LPS treatment. Apoptosis induced by LPS treatment was suppressed in the lungs of Chop-knockout mice. Overexpression of CHOP induced apoptosis in a lung cancer-derived cell line. These results suggest that the ER stress pathway, involving CHOP, is activated and plays a role in the pathogenesis of septic shock lung.
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Apoptosis/fisiología , Retículo Endoplásmico/metabolismo , Pulmón/metabolismo , Choque Séptico/metabolismo , Factor de Transcripción CHOP/metabolismo , Factor de Transcripción Activador 4/biosíntesis , Animales , Proteínas de Unión al ADN/biosíntesis , Modelos Animales de Enfermedad , Humanos , Lipopolisacáridos/farmacología , Pulmón/efectos de los fármacos , Ratones , Ratones Noqueados , Proteínas Nucleares/biosíntesis , ARN Mensajero/biosíntesis , ARN Mensajero/efectos de los fármacos , Factores de Transcripción del Factor Regulador X , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción CHOP/genética , Factores de Transcripción , Células Tumorales Cultivadas , Proteína 1 de Unión a la X-BoxRESUMEN
BACKGROUND/AIMS: There is little or no controversy about the increased oxidative stress of hemodialysis (HD) patients. Several reports show that the activity of superoxide dismutase (SOD), one of the major endogenous antioxidant enzymes, in plasma is elevated among HD patients. It is still unclear, however, whether this elevation is due to the promotion of SOD production or a decrease in renal excretion of SOD. This study was designed to investigate the cause of the SOD activation in HD patients, and we examined the expression of SOD mRNA levels in leukocytes of patients with chronic renal failure. METHODS: The total plasma SOD activity was determined by the nitroblue tetrazolium method, plasma SOD contents by ELISA, and SOD mRNA levels in leukocytes by RT-PCR. RESULTS: Our results demonstrated that contents and mRNA levels of Cu/Zn SOD in HD patients are 4.4 times and 2.0 times, respectively, as large as those in healthy controls. Furthermore, in contrast to nondialyzed chronic renal failure patients, we observed higher concentrations of Cu/Zn SOD in plasma and a more enhanced mRNA expression of Cu/Zn SOD in leukocytes of HD patients. CONCLUSION: Increased Cu/Zn SOD mRNA reflects enhanced antioxidant capacity of leukocytes and can be a promising oxidative stress marker in HD patients.
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Estrés Oxidativo , ARN Mensajero/biosíntesis , Diálisis Renal , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Femenino , Humanos , Leucocitos/enzimología , Masculino , Persona de Mediana Edad , Superóxido Dismutasa/sangreRESUMEN
PURPOSE: To understand the role of chondroitin sulfate proteoglycans during the development of rat cornea, expression of chondroitin sulfate and versican (PG-M) was studied. METHODS: Chondroitin sulfate and keratan sulfate in rat cornea were analyzed by immunohistochemical techniques. Reverse transcription polymerase chain reaction (RT-PCR) for chondroitin sulfate proteoglycans was performed. Versican expression was studied by RT-PCR, immunohistochemical, and dot blot analyses. Expression of hyaluronan was evaluated histochemically using biotinylated hyaluronan binding protein. RESULTS: Chondroitin sulfate was abundant in rat cornea at postnatal day 1 (P1) and became undetectable at P14. RT-PCR analysis showed that versican mRNA was highly expressed at P1 but was little expressed at P42. mRNAs for other chondroitin sulfate proteoglycans including biglycan, aggrecan, and decorin did not change much between P1 and P42. Expression for all versican splicing isoforms (V0-V3) was detectable from P1 through P14 but was undetectable after P21. mRNA for V0, the largest form with many chondroitin sulfate binding sites, decreased markedly in early stages from P1 to P14, whereas mRNA for V3, the shortest form with no chondroitin sulfate binding site, increased. mRNAs for middle-sized forms, V1 and V2, remained little changed during these periods. Immunohistochemical and dot blot analyses showed that versican is highly expressed at early stages of development and little expressed at adulthood. Similarly, hyaluronan, a versican-bound glycosaminoglycan, was highly expressed at early stages and little expressed at adulthood. CONCLUSIONS: Versican and hyaluronan, which can form a large molecular complex, may play an important role in the early phase of corneal development.